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Millipore novablue
Novablue, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 98 stars, based on 7 article reviews
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novablue - by Bioz Stars, 2020-01
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Clone Assay:

Article Title: CRISPR-spacer integration reporter plasmids reveal distinct genuine acquisition specificities among CRISPR-Cas I-E variants of Escherichia coli
Article Snippet: DNA amplicons to be cloned in plasmids other than linear pCR®2.1 vector ( ) were obtained with the Expand High Fidelity PCR System (Roche). .. Following ligation, pCR2.1 and pCDF-1b-derived plasmids were transformed in TOP10 (Invitrogen) or NovaBlue (Novagen) strains, respectively, and constructions were verified by sequencing with primers T7 (pCR2.1 and pCDF-1b) and M13R (pCR2.1) or T7t (pCDF-1b).

Article Title: Probing the Substrate Specificity and Protein-Protein Interactions of the E. coli Fatty Acid Dehydratase, FabA
Article Snippet: The FabA gene was cloned into the pET28b expression vector and used as the DNA template for site-directed mutagenesis with the mutagenic primer listed in . .. Digested PCR products were transformed into NovaBlue (EMD Millipore).

Article Title: Membrane integration of an essential β-barrel protein prerequires burial of an extracellular loop
Article Snippet: .. NovaBlue (Novagen) and BL21(DE3) (Novagen) strains were used for expression and cloning procedures. .. Proteinase K (Omega Bio-Tek) was dissolved in water and used directly in the protease digestion experiments.

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: .. The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained. .. To isolated purified protein, a Ni-NTA Fast Kit (Qiagen) was employed per the manufacturer’s instructions with the AmyA protein being present in the soluble fraction.

Article Title: Roles of Cysteine Proteases Cwp84 and Cwp13 in Biogenesis of the Cell Wall of Clostridium difficile ▿ ▿ †
Article Snippet: .. E. coli strains Top10 (Invitrogen) and NovaBlue (Novagen) were used routinely for cloning and propagation of plasmids. ..

Article Title: Association of Mycobacterium Proteins with Lipid Droplets
Article Snippet: .. Escherichia coli strains DH5α, Top10, (Thermo Fisher Scientific, Waltham, MA), and NovaBlue (MilliporeSigma, Burlington, MA) were used for cloning. .. Protein purification was carried out in E. coil strain BL21(DE3) containing plasmid pLysS.

Article Title: Detection and Assignment of Mutations and Minihaplotypes in Human DNA Using Peptide Mass Signature Genotyping (PMSG): Application to the HumanRDS/Peripherin Gene
Article Snippet: Paragraph title: Cloning, Transformation, and Expression ... Ligation products were transformed into NovaBlue(DE3) competent cells (Novagen) and outgrown in SOC for 1 h at 37°C with shaking at 300 rpm.

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: .. One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors. .. Recombinant plasmid DNA was purified with a Qiaprep kit (Qiagen).

Amplification:

Article Title: VE-statin/egfl7 Expression in Endothelial Cells Is Regulated by a Distal Enhancer and a Proximal Promoter under the Direct Control of Erg and GATA-2
Article Snippet: Region D. A similar scheme was applied except that the first amplification reactions were performed using −252/+38Luc (30.1fmoles) as template and the ScanD_n_fwd/GLprimer3 and RVprimer2/ScanD_n_rev primers, respectively ( ) using High Fidelity PCR Master Mix (Roche) in the supplied buffer and 5% dimethylsulfoxide. .. Vectors were transformed and then produced in Novablue (Novagen) or TG1 bacteria and purified in endotoxin-low conditions (PureLink™ HiPure Plasmid, Invitrogen).

Filtration:

Article Title: Structures of KcsA in Complex with Symmetrical Quaternary Ammonium Compounds Reveal a Hydrophobic Binding Site
Article Snippet: Briefly, the KcsA-L90C gene in pQE60 (Qiagen) was transformed into Novablue (Novagen) cells, and the cells were grown to logarithmic phase in Luria broth, induced with IPTG, and cultured. .. Thirty-five residues of KcsA were removed with chymotrypsin, and the truncated channel was further purified by gel filtration on a Superdex 200 column (GE Healthcare).

Synthesized:

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: Oligonucleotide primers were synthesized by Integrated DNA Technologies (Coralville, IA). .. One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors.

Construct:

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: .. The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained. .. To isolated purified protein, a Ni-NTA Fast Kit (Qiagen) was employed per the manufacturer’s instructions with the AmyA protein being present in the soluble fraction.

Incubation:

Article Title: Association of Mycobacterium Proteins with Lipid Droplets
Article Snippet: Escherichia coli strains DH5α, Top10, (Thermo Fisher Scientific, Waltham, MA), and NovaBlue (MilliporeSigma, Burlington, MA) were used for cloning. .. All E. coli strains were incubated at 37°C in Luria-Bertani (LB) medium with 100 μg/ml ampicillin, 50 μg/ml kanamycin, 150 μg/ml hygromycin B, and/or 25 μg/ml chloramphenicol for selection, as necessary.

Article Title: Detection and Assignment of Mutations and Minihaplotypes in Human DNA Using Peptide Mass Signature Genotyping (PMSG): Application to the HumanRDS/Peripherin Gene
Article Snippet: Ligation products were transformed into NovaBlue(DE3) competent cells (Novagen) and outgrown in SOC for 1 h at 37°C with shaking at 300 rpm. .. The following morning, fresh media was added and cells were incubated for an additional 3 h before IPTG to 1 mM was added and cultures were incubated for one additional hour.

Activity Assay:

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: Paragraph title: Overexpression and purification of AmyA and assessment of α-glucan degrading activity ... The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained.

Infection:

Article Title: The Ability To Replicate in Macrophages Is Conserved between Yersinia pestis and Yersinia pseudotuberculosis
Article Snippet: Cultures were grown in broth with aeration for 20 h at 26°C to prepare bacteria for infection assays. .. The Escherichia coli strains used, NovaBlue (Novagen), DH5α , and S17-1 ( ) lysogenized with λpir , were grown in LB broth or on LB agar plates at 37°C.

Expressing:

Article Title: Probing the Substrate Specificity and Protein-Protein Interactions of the E. coli Fatty Acid Dehydratase, FabA
Article Snippet: The FabA gene was cloned into the pET28b expression vector and used as the DNA template for site-directed mutagenesis with the mutagenic primer listed in . .. Digested PCR products were transformed into NovaBlue (EMD Millipore).

Article Title: Membrane integration of an essential β-barrel protein prerequires burial of an extracellular loop
Article Snippet: .. NovaBlue (Novagen) and BL21(DE3) (Novagen) strains were used for expression and cloning procedures. .. Proteinase K (Omega Bio-Tek) was dissolved in water and used directly in the protease digestion experiments.

Article Title: Structures of KcsA in Complex with Symmetrical Quaternary Ammonium Compounds Reveal a Hydrophobic Binding Site
Article Snippet: Paragraph title: Protein Expression and Purification ... Briefly, the KcsA-L90C gene in pQE60 (Qiagen) was transformed into Novablue (Novagen) cells, and the cells were grown to logarithmic phase in Luria broth, induced with IPTG, and cultured.

Article Title: An ATP-Grasp Ligase Involved in the Last Biosynthetic Step of the Iminomycosporine Shinorine in Nostoc punctiforme ATCC 29133 ▿
Article Snippet: Escherichia coli DH5α or NovaBlue (Novagen/EMD Biosciences, San Diego, CA) competent cells were used for standard subcloning. .. E. coli BL21(DE3) (Novagen) was used for gene expression and for in vivo bioconversion.

Article Title: Detection and Assignment of Mutations and Minihaplotypes in Human DNA Using Peptide Mass Signature Genotyping (PMSG): Application to the HumanRDS/Peripherin Gene
Article Snippet: Paragraph title: Cloning, Transformation, and Expression ... Ligation products were transformed into NovaBlue(DE3) competent cells (Novagen) and outgrown in SOC for 1 h at 37°C with shaking at 300 rpm.

Bradford Assay:

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained. .. The amount of AmyA present was determined using a Bradford assay (Bio-Rad).

Modification:

Article Title: Detection and Assignment of Mutations and Minihaplotypes in Human DNA Using Peptide Mass Signature Genotyping (PMSG): Application to the HumanRDS/Peripherin Gene
Article Snippet: WZ4 is a modified pET24d+ plasmid (Novagen). .. Ligation products were transformed into NovaBlue(DE3) competent cells (Novagen) and outgrown in SOC for 1 h at 37°C with shaking at 300 rpm.

Western Blot:

Article Title: Membrane integration of an essential β-barrel protein prerequires burial of an extracellular loop
Article Snippet: FLAG-BamA was detected on Western blots with monoclonal anti-FLAG M2-peroxidase (HRP) mouse antibody from Sigma-Aldrich. .. NovaBlue (Novagen) and BL21(DE3) (Novagen) strains were used for expression and cloning procedures.

Transformation Assay:

Article Title: CRISPR-spacer integration reporter plasmids reveal distinct genuine acquisition specificities among CRISPR-Cas I-E variants of Escherichia coli
Article Snippet: .. Following ligation, pCR2.1 and pCDF-1b-derived plasmids were transformed in TOP10 (Invitrogen) or NovaBlue (Novagen) strains, respectively, and constructions were verified by sequencing with primers T7 (pCR2.1 and pCDF-1b) and M13R (pCR2.1) or T7t (pCDF-1b). .. Detection of CRISPR, leader and PAM sequences Leaders of ECOR strains were determined by sequencing PCR amplification products as described by Diez-Villaseñor et al. CRISPR and leaders in available E. coli genomes were identified with the CRISPRFinder program ( www.crispr.u-psud.fr/Server/ ).

Article Title: Probing the Substrate Specificity and Protein-Protein Interactions of the E. coli Fatty Acid Dehydratase, FabA
Article Snippet: .. Digested PCR products were transformed into NovaBlue (EMD Millipore). .. Single colonies were isolated and their plasmids were extracted using Qiagen Minipreps.

Article Title: Structures of KcsA in Complex with Symmetrical Quaternary Ammonium Compounds Reveal a Hydrophobic Binding Site
Article Snippet: .. Briefly, the KcsA-L90C gene in pQE60 (Qiagen) was transformed into Novablue (Novagen) cells, and the cells were grown to logarithmic phase in Luria broth, induced with IPTG, and cultured. .. After cell membranes were disrupted by sonication, KcsA was solubilized with decyl maltoside (DM) and purified on a cobalt affinity column (Talon, Clontech).

Article Title: VE-statin/egfl7 Expression in Endothelial Cells Is Regulated by a Distal Enhancer and a Proximal Promoter under the Direct Control of Erg and GATA-2
Article Snippet: .. Vectors were transformed and then produced in Novablue (Novagen) or TG1 bacteria and purified in endotoxin-low conditions (PureLink™ HiPure Plasmid, Invitrogen). ..

Article Title: Detection and Assignment of Mutations and Minihaplotypes in Human DNA Using Peptide Mass Signature Genotyping (PMSG): Application to the HumanRDS/Peripherin Gene
Article Snippet: .. Ligation products were transformed into NovaBlue(DE3) competent cells (Novagen) and outgrown in SOC for 1 h at 37°C with shaking at 300 rpm. ..

Over Expression:

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: Paragraph title: Overexpression and purification of AmyA and assessment of α-glucan degrading activity ... The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained.

Derivative Assay:

Article Title: Global Gene Expression Profiling of Yersinia pestis Replicating inside Macrophages Reveals the Roles of a Putative Stress-Induced Operon in Regulating Type III Secretion and Intracellular Cell Division ▿ Replicating inside Macrophages Reveals the Roles of a Putative Stress-Induced Operon in Regulating Type III Secretion and Intracellular Cell Division ▿ †
Article Snippet: The Y. pestis strains used in this study (Table ) are derived from strain KIM (molecular group 2.MED [ ]). .. Escherichia coli strains S17-1 and NovaBlue (Novagen) were grown on LB agar or LB broth at 37°C.

Ligation:

Article Title: CRISPR-spacer integration reporter plasmids reveal distinct genuine acquisition specificities among CRISPR-Cas I-E variants of Escherichia coli
Article Snippet: .. Following ligation, pCR2.1 and pCDF-1b-derived plasmids were transformed in TOP10 (Invitrogen) or NovaBlue (Novagen) strains, respectively, and constructions were verified by sequencing with primers T7 (pCR2.1 and pCDF-1b) and M13R (pCR2.1) or T7t (pCDF-1b). .. Detection of CRISPR, leader and PAM sequences Leaders of ECOR strains were determined by sequencing PCR amplification products as described by Diez-Villaseñor et al. CRISPR and leaders in available E. coli genomes were identified with the CRISPRFinder program ( www.crispr.u-psud.fr/Server/ ).

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: .. The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained. .. To isolated purified protein, a Ni-NTA Fast Kit (Qiagen) was employed per the manufacturer’s instructions with the AmyA protein being present in the soluble fraction.

Article Title: Detection and Assignment of Mutations and Minihaplotypes in Human DNA Using Peptide Mass Signature Genotyping (PMSG): Application to the HumanRDS/Peripherin Gene
Article Snippet: .. Ligation products were transformed into NovaBlue(DE3) competent cells (Novagen) and outgrown in SOC for 1 h at 37°C with shaking at 300 rpm. ..

Cell Culture:

Article Title: Structures of KcsA in Complex with Symmetrical Quaternary Ammonium Compounds Reveal a Hydrophobic Binding Site
Article Snippet: .. Briefly, the KcsA-L90C gene in pQE60 (Qiagen) was transformed into Novablue (Novagen) cells, and the cells were grown to logarithmic phase in Luria broth, induced with IPTG, and cultured. .. After cell membranes were disrupted by sonication, KcsA was solubilized with decyl maltoside (DM) and purified on a cobalt affinity column (Talon, Clontech).

Article Title: Association of Mycobacterium Proteins with Lipid Droplets
Article Snippet: Escherichia coli strains DH5α, Top10, (Thermo Fisher Scientific, Waltham, MA), and NovaBlue (MilliporeSigma, Burlington, MA) were used for cloning. .. Rhodococcus jostii strain RHA1 was kindly provided by Lindsay Eltis (University of British Columbia, Canada) and was cultured at 30°C in LB medium.

DNA Sequencing:

Article Title: Probing the Substrate Specificity and Protein-Protein Interactions of the E. coli Fatty Acid Dehydratase, FabA
Article Snippet: Digested PCR products were transformed into NovaBlue (EMD Millipore). .. Site-directed mutagenesis was verified via DNA sequencing (IDT).

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors. .. DNA sequencing was performed at the Molecular Biology Core Facilities of the Dana Farber Cancer Institute (Boston, MA).

Polymerase Chain Reaction:

Article Title: CRISPR-spacer integration reporter plasmids reveal distinct genuine acquisition specificities among CRISPR-Cas I-E variants of Escherichia coli
Article Snippet: PCR products were purified and afterwards digested with Thermo Scientific enzymes according to Double Digest web tool recommendations ( www.thermoscientificbio.com/webtools/doubledigest/ ). .. Following ligation, pCR2.1 and pCDF-1b-derived plasmids were transformed in TOP10 (Invitrogen) or NovaBlue (Novagen) strains, respectively, and constructions were verified by sequencing with primers T7 (pCR2.1 and pCDF-1b) and M13R (pCR2.1) or T7t (pCDF-1b).

Article Title: Probing the Substrate Specificity and Protein-Protein Interactions of the E. coli Fatty Acid Dehydratase, FabA
Article Snippet: .. Digested PCR products were transformed into NovaBlue (EMD Millipore). .. Single colonies were isolated and their plasmids were extracted using Qiagen Minipreps.

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: The resultant PCR product was digested with Xho I and EcoR 1 and ligated into the pBAD-hisA plasmid (Invitrogen) that had also been digested with the same enzymes. .. The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained.

Article Title: VE-statin/egfl7 Expression in Endothelial Cells Is Regulated by a Distal Enhancer and a Proximal Promoter under the Direct Control of Erg and GATA-2
Article Snippet: Thermal conditions were 95°C/5min, 33 cycles [94°C/30s, 55°C/30s, 72°C/50s], 72°C/7 min. PCR products were digested and inserted into the NheI/HindIII site of pGL3basic. .. Vectors were transformed and then produced in Novablue (Novagen) or TG1 bacteria and purified in endotoxin-low conditions (PureLink™ HiPure Plasmid, Invitrogen).

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: Polymerase chain reaction (PCR) was performed with Phusion High-Fidelity PCR Mastermix (New England Biolabs). .. One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors.

Sonication:

Article Title: Structures of KcsA in Complex with Symmetrical Quaternary Ammonium Compounds Reveal a Hydrophobic Binding Site
Article Snippet: Briefly, the KcsA-L90C gene in pQE60 (Qiagen) was transformed into Novablue (Novagen) cells, and the cells were grown to logarithmic phase in Luria broth, induced with IPTG, and cultured. .. After cell membranes were disrupted by sonication, KcsA was solubilized with decyl maltoside (DM) and purified on a cobalt affinity column (Talon, Clontech).

Recombinant:

Article Title: CRISPR-spacer integration reporter plasmids reveal distinct genuine acquisition specificities among CRISPR-Cas I-E variants of Escherichia coli
Article Snippet: For cloning into the 3′-T overhangs of the linearized pCR®2.1 vector, PCR products were obtained with recombinant Taq polymerase (Invitrogen). .. Following ligation, pCR2.1 and pCDF-1b-derived plasmids were transformed in TOP10 (Invitrogen) or NovaBlue (Novagen) strains, respectively, and constructions were verified by sequencing with primers T7 (pCR2.1 and pCDF-1b) and M13R (pCR2.1) or T7t (pCDF-1b).

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors. .. Recombinant plasmid DNA was purified with a Qiaprep kit (Qiagen).

Nucleic Acid Electrophoresis:

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors. .. Nickel-nitrilotriacetic acid-agarose (Ni-NTA) superflow resin and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels were purchased from Qiagen and Biorad, respectively.

In Vivo:

Article Title: An ATP-Grasp Ligase Involved in the Last Biosynthetic Step of the Iminomycosporine Shinorine in Nostoc punctiforme ATCC 29133 ▿
Article Snippet: Escherichia coli DH5α or NovaBlue (Novagen/EMD Biosciences, San Diego, CA) competent cells were used for standard subcloning. .. E. coli BL21(DE3) (Novagen) was used for gene expression and for in vivo bioconversion.

Mutagenesis:

Article Title: Probing the Substrate Specificity and Protein-Protein Interactions of the E. coli Fatty Acid Dehydratase, FabA
Article Snippet: Paragraph title: Site-directed Mutagenesis ... Digested PCR products were transformed into NovaBlue (EMD Millipore).

Isolation:

Article Title: Probing the Substrate Specificity and Protein-Protein Interactions of the E. coli Fatty Acid Dehydratase, FabA
Article Snippet: Digested PCR products were transformed into NovaBlue (EMD Millipore). .. Single colonies were isolated and their plasmids were extracted using Qiagen Minipreps.

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained. .. To isolated purified protein, a Ni-NTA Fast Kit (Qiagen) was employed per the manufacturer’s instructions with the AmyA protein being present in the soluble fraction.

Subcloning:

Article Title: An ATP-Grasp Ligase Involved in the Last Biosynthetic Step of the Iminomycosporine Shinorine in Nostoc punctiforme ATCC 29133 ▿
Article Snippet: .. Escherichia coli DH5α or NovaBlue (Novagen/EMD Biosciences, San Diego, CA) competent cells were used for standard subcloning. .. E. coli BL21(DE3) (Novagen) was used for gene expression and for in vivo bioconversion.

Purification:

Article Title: Mutations in RNA Polymerase Bridge Helix and Switch Regions Affect Active-Site Networks and Transcript-Assisted Hydrolysis
Article Snippet: .. The reconstituted RNAP mutants were over-expressed in NovaBlue (Novagen) and purified as previously described . .. E . coli PspF1 -275 (the AAA+ domain, residues 1–275) and Klebsiella pneumoniae σ54 were over-expressed in BL21(DE3) and purified as previously described .

Article Title: CRISPR-spacer integration reporter plasmids reveal distinct genuine acquisition specificities among CRISPR-Cas I-E variants of Escherichia coli
Article Snippet: PCR products were purified and afterwards digested with Thermo Scientific enzymes according to Double Digest web tool recommendations ( www.thermoscientificbio.com/webtools/doubledigest/ ). .. Following ligation, pCR2.1 and pCDF-1b-derived plasmids were transformed in TOP10 (Invitrogen) or NovaBlue (Novagen) strains, respectively, and constructions were verified by sequencing with primers T7 (pCR2.1 and pCDF-1b) and M13R (pCR2.1) or T7t (pCDF-1b).

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: Paragraph title: Overexpression and purification of AmyA and assessment of α-glucan degrading activity ... The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained.

Article Title: Structures of KcsA in Complex with Symmetrical Quaternary Ammonium Compounds Reveal a Hydrophobic Binding Site
Article Snippet: Paragraph title: Protein Expression and Purification ... Briefly, the KcsA-L90C gene in pQE60 (Qiagen) was transformed into Novablue (Novagen) cells, and the cells were grown to logarithmic phase in Luria broth, induced with IPTG, and cultured.

Article Title: VE-statin/egfl7 Expression in Endothelial Cells Is Regulated by a Distal Enhancer and a Proximal Promoter under the Direct Control of Erg and GATA-2
Article Snippet: .. Vectors were transformed and then produced in Novablue (Novagen) or TG1 bacteria and purified in endotoxin-low conditions (PureLink™ HiPure Plasmid, Invitrogen). ..

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors. .. Recombinant plasmid DNA was purified with a Qiaprep kit (Qiagen).

Protein Purification:

Article Title: Association of Mycobacterium Proteins with Lipid Droplets
Article Snippet: Escherichia coli strains DH5α, Top10, (Thermo Fisher Scientific, Waltham, MA), and NovaBlue (MilliporeSigma, Burlington, MA) were used for cloning. .. Protein purification was carried out in E. coil strain BL21(DE3) containing plasmid pLysS.

Sequencing:

Article Title: CRISPR-spacer integration reporter plasmids reveal distinct genuine acquisition specificities among CRISPR-Cas I-E variants of Escherichia coli
Article Snippet: .. Following ligation, pCR2.1 and pCDF-1b-derived plasmids were transformed in TOP10 (Invitrogen) or NovaBlue (Novagen) strains, respectively, and constructions were verified by sequencing with primers T7 (pCR2.1 and pCDF-1b) and M13R (pCR2.1) or T7t (pCDF-1b). .. Detection of CRISPR, leader and PAM sequences Leaders of ECOR strains were determined by sequencing PCR amplification products as described by Diez-Villaseñor et al. CRISPR and leaders in available E. coli genomes were identified with the CRISPRFinder program ( www.crispr.u-psud.fr/Server/ ).

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: .. The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained. .. To isolated purified protein, a Ni-NTA Fast Kit (Qiagen) was employed per the manufacturer’s instructions with the AmyA protein being present in the soluble fraction.

Article Title: Detection and Assignment of Mutations and Minihaplotypes in Human DNA Using Peptide Mass Signature Genotyping (PMSG): Application to the HumanRDS/Peripherin Gene
Article Snippet: The coding sequence for the universal epitope ( ) was placed immediately downstream of the initial ATG. .. Ligation products were transformed into NovaBlue(DE3) competent cells (Novagen) and outgrown in SOC for 1 h at 37°C with shaking at 300 rpm.

Affinity Column:

Article Title: Mutations in RNA Polymerase Bridge Helix and Switch Regions Affect Active-Site Networks and Transcript-Assisted Hydrolysis
Article Snippet: The reconstituted RNAP mutants were over-expressed in NovaBlue (Novagen) and purified as previously described . .. E . coli σ70 was over-expressed in BL21(DE3) and purified via a nickel affinity column followed by a heparin chelating step.

Article Title: Structures of KcsA in Complex with Symmetrical Quaternary Ammonium Compounds Reveal a Hydrophobic Binding Site
Article Snippet: Briefly, the KcsA-L90C gene in pQE60 (Qiagen) was transformed into Novablue (Novagen) cells, and the cells were grown to logarithmic phase in Luria broth, induced with IPTG, and cultured. .. After cell membranes were disrupted by sonication, KcsA was solubilized with decyl maltoside (DM) and purified on a cobalt affinity column (Talon, Clontech).

IA:

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: Oligonucleotide primers were synthesized by Integrated DNA Technologies (Coralville, IA). .. One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors.

SDS Page:

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors. .. Nickel-nitrilotriacetic acid-agarose (Ni-NTA) superflow resin and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels were purchased from Qiagen and Biorad, respectively.

Plasmid Preparation:

Article Title: CRISPR-spacer integration reporter plasmids reveal distinct genuine acquisition specificities among CRISPR-Cas I-E variants of Escherichia coli
Article Snippet: DNA amplicons to be cloned in plasmids other than linear pCR®2.1 vector ( ) were obtained with the Expand High Fidelity PCR System (Roche). .. Following ligation, pCR2.1 and pCDF-1b-derived plasmids were transformed in TOP10 (Invitrogen) or NovaBlue (Novagen) strains, respectively, and constructions were verified by sequencing with primers T7 (pCR2.1 and pCDF-1b) and M13R (pCR2.1) or T7t (pCDF-1b).

Article Title: Probing the Substrate Specificity and Protein-Protein Interactions of the E. coli Fatty Acid Dehydratase, FabA
Article Snippet: The FabA gene was cloned into the pET28b expression vector and used as the DNA template for site-directed mutagenesis with the mutagenic primer listed in . .. Digested PCR products were transformed into NovaBlue (EMD Millipore).

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: The resultant PCR product was digested with Xho I and EcoR 1 and ligated into the pBAD-hisA plasmid (Invitrogen) that had also been digested with the same enzymes. .. The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained.

Article Title: VE-statin/egfl7 Expression in Endothelial Cells Is Regulated by a Distal Enhancer and a Proximal Promoter under the Direct Control of Erg and GATA-2
Article Snippet: .. Vectors were transformed and then produced in Novablue (Novagen) or TG1 bacteria and purified in endotoxin-low conditions (PureLink™ HiPure Plasmid, Invitrogen). ..

Article Title: Recombinant E.coli prototype strains for in vivo glycorandomization
Article Snippet: NovaBlue was from Novagen. .. Plasmid pET28/OleD was a generous gift from Prof Hung-Wen Liu (University of Texas-Austin, Austin, USA) and pET28a was from Novagen.

Article Title: Association of Mycobacterium Proteins with Lipid Droplets
Article Snippet: Escherichia coli strains DH5α, Top10, (Thermo Fisher Scientific, Waltham, MA), and NovaBlue (MilliporeSigma, Burlington, MA) were used for cloning. .. Protein purification was carried out in E. coil strain BL21(DE3) containing plasmid pLysS.

Article Title: Detection and Assignment of Mutations and Minihaplotypes in Human DNA Using Peptide Mass Signature Genotyping (PMSG): Application to the HumanRDS/Peripherin Gene
Article Snippet: The plasmid carries the lacIq gene and the npt1 gene, which confers resistance to kanamycin. .. Ligation products were transformed into NovaBlue(DE3) competent cells (Novagen) and outgrown in SOC for 1 h at 37°C with shaking at 300 rpm.

Article Title: The NRPS Enzyme DdaD Tethers Nβ-fumaramoyl-DAP for Fe(II)/?-ketoglutarate-Dependent Epoxidation by DdaC During Dapdiamide Antibiotic Biosynthesis
Article Snippet: One Shot Chemically Competent TOP10 E. coli (Invitrogen) and NovaBlue(DE3) (Novagen) were used for routine cloning and propagation of DNA vectors. .. Recombinant plasmid DNA was purified with a Qiaprep kit (Qiagen).

Selection:

Article Title: Contribution of AmyA, an extracellular α-glucan degrading enzyme, to group A streptococcal host-pathogen interaction
Article Snippet: .. The ligation product, called pBAD-amyA, was cloned into Novablue (Novagen) and then BL21 (Novagen) competent Escherichia coli cells with selection via ampicillin resistance. pBAD-amyA was analyzed via sequencing to ensure that the correct construct had been obtained. .. To isolated purified protein, a Ni-NTA Fast Kit (Qiagen) was employed per the manufacturer’s instructions with the AmyA protein being present in the soluble fraction.

Article Title: Roles of Cysteine Proteases Cwp84 and Cwp13 in Biogenesis of the Cell Wall of Clostridium difficile ▿ ▿ †
Article Snippet: E. coli strains Top10 (Invitrogen) and NovaBlue (Novagen) were used routinely for cloning and propagation of plasmids. .. E. coli strains were routinely grown in LB broth ( ) or on LB agar supplemented with appropriate antibiotics: carbenicillin at 50 μg/ml for the selection of derivatives of pUC19 and chloramphenicol at 12.5 μg/ml for the selection of derivatives of pMTL007 or pMTL960.

Article Title: Association of Mycobacterium Proteins with Lipid Droplets
Article Snippet: Escherichia coli strains DH5α, Top10, (Thermo Fisher Scientific, Waltham, MA), and NovaBlue (MilliporeSigma, Burlington, MA) were used for cloning. .. All E. coli strains were incubated at 37°C in Luria-Bertani (LB) medium with 100 μg/ml ampicillin, 50 μg/ml kanamycin, 150 μg/ml hygromycin B, and/or 25 μg/ml chloramphenicol for selection, as necessary.

Produced:

Article Title: VE-statin/egfl7 Expression in Endothelial Cells Is Regulated by a Distal Enhancer and a Proximal Promoter under the Direct Control of Erg and GATA-2
Article Snippet: .. Vectors were transformed and then produced in Novablue (Novagen) or TG1 bacteria and purified in endotoxin-low conditions (PureLink™ HiPure Plasmid, Invitrogen). ..

Concentration Assay:

Article Title: Probing the Substrate Specificity and Protein-Protein Interactions of the E. coli Fatty Acid Dehydratase, FabA
Article Snippet: Polymerase chain reactions (PCR) were performed using a combination of varying DMSO and magnesium concentration and “step down” annealing temperatures. .. Digested PCR products were transformed into NovaBlue (EMD Millipore).

Article Title: Membrane integration of an essential β-barrel protein prerequires burial of an extracellular loop
Article Snippet: Kanamycin (kan) and carbenicillin (carb) were used at a final concentration of 50 μg/mL, chloramphenicol (cam) was used at a final concentration of 30 μg/mL. .. NovaBlue (Novagen) and BL21(DE3) (Novagen) strains were used for expression and cloning procedures.

Chick Chorioallantoic Membrane Assay:

Article Title: Membrane integration of an essential β-barrel protein prerequires burial of an extracellular loop
Article Snippet: Kanamycin (kan) and carbenicillin (carb) were used at a final concentration of 50 μg/mL, chloramphenicol (cam) was used at a final concentration of 30 μg/mL. .. NovaBlue (Novagen) and BL21(DE3) (Novagen) strains were used for expression and cloning procedures.

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